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Chemical shift assignments and the secondary structure of the Est3 telomerase subunit in the yeast Hansenula polymorpha

Biomolecular NMR Assignments volume 12, pages 57–62 (2018)Cite this article

Abstract

Telomerase is a multisubunit ribonucleoprotein enzyme that is essential for continuous cellular proliferation. A key role of telomerase in cancer and ageing makes it a promising target for the development of cancer therapies and treatments of other age-associated diseases, since telomerase allows unlimited proliferation potential of cells in the majority of cancer types. However, the structure and molecular mechanism of telomerase action are still poorly understood. In budding yeast, telomerase consists of the catalytic subunit, the telomerase reverse transcriptase or Est2 protein, telomerase RNA (TLC1) and two regulatory subunits, Est1 and Est3. Each of the four subunits is essential for in vivo telomerase function. Est3 interacts directly with Est1 and Est2, and stimulates Est2 catalytic activity. However, the exact role of the Est3 protein in telomerase function is still unknown. Determination of the structure, dynamic and functional properties of Est3 can bring new insights into the molecular mechanism of telomerase activity. Here we report nearly complete 1H, 13C and 15N resonance assignments of Est3 from the yeast Hansenula polymorpha. Analysis of the assigned chemical shifts allowed us to identify the protein’s secondary structure and backbone dynamic properties. Structure-based sequence alignment revealed similarities in the structural organization of yeast Est3 and mammalian TPP1 proteins.

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Abbreviations

IPTG:

Isopropyl-thio-β-d-galactoside

Est1:

Telomere elongation protein

Est2:

Telomerase reverse transcriptase subunit

Est3:

Telomere replication protein

hpEst3:

Hansenula polymorpha Est3 protein

scEst3:

Saccharomyces cerevisiae Est3 protein

TEN:

Telomerase N-terminal domain

TER:

Telomerase RNA

TERT:

Telomerase reverse transcriptase subunit

TEV:

Tobacco etch virus nuclear-inclusion-a endopeptidase

TPPI:

Telomere-binding protein, component of the six-membered shelterin complex

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Acknowledgements

The Russian Science Foundation (Grant 14-14-00598) supported the NMR studies. The Russian Foundation for Basic Research (Grant 15-54-74005 EMBL_a) supported protein cloning, expression and purification. The authors are grateful for the opportunity to use the NMR facilities acquired with the support from the Russian Government Program of Competitive Growth of Kazan Federal University, among the world’s leading academic centers.

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Authors and Affiliations

  1. Center for Magnetic Tomography and Spectroscopy, Faculty of Fundamental Medicine, M.V. Lomonosov Moscow State University, Moscow, Russia, 119991

    Sofia S. Mariasina & Vladimir I. Polshakov

  2. Department of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia, 119991

    Sofia S. Mariasina, Elena V. Rodina, Alexander N. Malyavko, Maria I. Zvereva & Olga A. Dontsova

  3. NMR Laboratory, Institute of Physics, Kazan Federal University, 18 Kremlevskaya, Kazan, Russia, 420008

    Sergey V. Efimov & Vladimir V. Klochkov

  4. A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Moscow, Russia, 119991

    Olga A. Petrova & Olga A. Dontsova

  5. Skolkovo Institute of Science and Technology, Moscow, Russia

    Alexander N. Malyavko & Olga A. Dontsova

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  1. Sofia S. Mariasina
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  2. Sergey V. Efimov
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  3. Olga A. Petrova
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  4. Elena V. Rodina
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  9. Vladimir I. Polshakov
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Correspondence to Vladimir I. Polshakov.

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Mariasina, S.S., Efimov, S.V., Petrova, O.A. et al. Chemical shift assignments and the secondary structure of the Est3 telomerase subunit in the yeast Hansenula polymorpha . Biomol NMR Assign 12, 57–62 (2018). https://doi.org/10.1007/s12104-017-9780-5

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Keywords

  • Telomerase
  • Protein NMR
  • Resonance assignment
  • Secondary structure